This document provides information on using the BioPAT Trace system to monitor glucose and lactate concentrations in bioreactor processes. It discusses the measurement principles, which use the enzymes glucose oxidase and lactate oxidase. Data is presented to demonstrate the system's linearity, precision, recovery, operational stability, and shelf life for glucose and lactate sensors. Consumables required for using the system are also listed.
Analysis of Phenolic Antioxidants in Edible Oil/Shortening Using the PerkinEl...PerkinElmer, Inc.
Phenolic antioxidants are commonly used in food to prevent the oxidation of oils. Oxidized oil and fats cause foul odor and rancidity in food products, which is a major cause for concern to the food industry. Globally, regulations vary, but current maximum allowable levels are as low as 100 μg/g (100 ppm). This application note presents a UHPLC method for the analysis of the ten most common phenolic antioxidants that may be found in such products.
Analysis of Sugars in Honey Using the PerkinElmer Altus HPLC System with RI D...PerkinElmer, Inc.
Honey consumption has grown significantly during the last few decades due to its high nutritional value and unique flavor. The price of natural bee honey is much higher than other sweeteners making it susceptible to adulteration with cheaper sweeteners, primarily sucrose. Besides lower levels of nonsugar ingredients, natural honey primarily consists of glucose and fructose and may contain low levels of sucrose and/or maltose. However, according to the international regulations, any commercially available “pure”-labeled honey products that are found to have in excess of 5% by weight of sucrose or maltose are considered to be adulterated. With the focus on possible honey adulteration, this application highlights the LC separation of various sugars found in honey and the analysis of these components in four store-bought honey samples. Method conditions and performance data, including linearity and repeatability, are presented.
Improving Galacto-Oligosaccharide Content in the Production of Lactose-Reduce...IJERA Editor
In a lactic fermentation process with probiotic microorganisms and the simultaneous addition of -
galactosidase, the reduction of lactose content and the formation of galacto-oligosaccharides were evaluated. The
fermentation was promoted by lactic culture containing two probiotic microorganisms, Bifidobacterium animalis
and Lactobacillus acidophilus, associated with the typical microorganisms of yogurt, Lactobacillus bulgaricus
and Streptococcus thermophilus. An enzymatic preparation containing -galactosidases from Kluyveromyces
lactis and Aspergillus niger was used. A central composite rotational design (CCRD) and a total of 10 assays (22
assays plus axial points and two replicates at the central point) were conducted in order to evaluate the effects of
enzyme concentration and the time of addition of the enzyme. Based on an experimental design, empirical
models for the final lactose concentration and GOS concentration were proposed. The following conditions were
established in order to maximize GOS concentration: enzyme concentration of 0.44 g/L and enzyme addition
after 90 min from the beginning of fermentation. In these conditions a ten-fold increase in GOS concentration
and a four-fold decrease in lactose concentration were observed in comparison with fermentation without
enzyme addition.
Analysis of Phenolic Antioxidants in Edible Oil/Shortening Using the PerkinEl...PerkinElmer, Inc.
Phenolic antioxidants are commonly used in food to prevent the oxidation of oils. Oxidized oil and fats cause foul odor and rancidity in food products, which is a major cause for concern to the food industry. Globally, regulations vary, but current maximum allowable levels are as low as 100 μg/g (100 ppm). This application note presents a UHPLC method for the analysis of the ten most common phenolic antioxidants that may be found in such products.
Analysis of Sugars in Honey Using the PerkinElmer Altus HPLC System with RI D...PerkinElmer, Inc.
Honey consumption has grown significantly during the last few decades due to its high nutritional value and unique flavor. The price of natural bee honey is much higher than other sweeteners making it susceptible to adulteration with cheaper sweeteners, primarily sucrose. Besides lower levels of nonsugar ingredients, natural honey primarily consists of glucose and fructose and may contain low levels of sucrose and/or maltose. However, according to the international regulations, any commercially available “pure”-labeled honey products that are found to have in excess of 5% by weight of sucrose or maltose are considered to be adulterated. With the focus on possible honey adulteration, this application highlights the LC separation of various sugars found in honey and the analysis of these components in four store-bought honey samples. Method conditions and performance data, including linearity and repeatability, are presented.
Improving Galacto-Oligosaccharide Content in the Production of Lactose-Reduce...IJERA Editor
In a lactic fermentation process with probiotic microorganisms and the simultaneous addition of -
galactosidase, the reduction of lactose content and the formation of galacto-oligosaccharides were evaluated. The
fermentation was promoted by lactic culture containing two probiotic microorganisms, Bifidobacterium animalis
and Lactobacillus acidophilus, associated with the typical microorganisms of yogurt, Lactobacillus bulgaricus
and Streptococcus thermophilus. An enzymatic preparation containing -galactosidases from Kluyveromyces
lactis and Aspergillus niger was used. A central composite rotational design (CCRD) and a total of 10 assays (22
assays plus axial points and two replicates at the central point) were conducted in order to evaluate the effects of
enzyme concentration and the time of addition of the enzyme. Based on an experimental design, empirical
models for the final lactose concentration and GOS concentration were proposed. The following conditions were
established in order to maximize GOS concentration: enzyme concentration of 0.44 g/L and enzyme addition
after 90 min from the beginning of fermentation. In these conditions a ten-fold increase in GOS concentration
and a four-fold decrease in lactose concentration were observed in comparison with fermentation without
enzyme addition.
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This work will describe measurements of a variety of toxic metals at low concentrations in fruit juices and fruit purees. Sample preparation and the effect on detection limits will be described. Graphite furnace atomic absorption (GFAA) and inductively coupled plasma mass spectrometry (ICP-MS) will be compared and an overall approach to analysis described.
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This presentation describes a new food testing solution that allows mass detection to be accessible for many of the routine analyses found within a food testing lab. The ACQUITY UPLC, QDa detector in combination with the Empower software solution is fit for purpose and is easy to use. You simply power on and you are ready to go
With the ACQUITY QDa detector many of the normal processes that are required for mass spectrometers (such as mass calibration and optimisation and manual adjustments that need to be made) have all been fully automated.
In this application note, the amount of sugar or carbohydrate in a soft drink was determined using a colorimetric method. The rapid measurement of the PDA (Photodiode Array) UV/Vis Spectrophotometer allows for the collection of accurate data from the time-dependent reaction. The calibration curve was automatically calculated using the Quantification mode of the UV Lab™ software.
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This work describes a simple and
direct dilution method for sample preparation, followed by
automated analysis using GFAAS. This method minimizes
sample preparation, and also reduces potential contamination
while still maintaining the speed of analysis.
Learn more about our solutions: http://bit.ly/1bXfnRZ
An overview of Pine Lake Laboratories capabilities involving oligonucleotides. Includes challenges, examples, method development, validation, and stability!
The Analysis of Baby Foods and Juices for Metals to Protect a Sensitive Popul...PerkinElmer, Inc.
This work will describe measurements of a variety of toxic metals at low concentrations in fruit juices and fruit purees. Sample preparation and the effect on detection limits will be described. Graphite furnace atomic absorption (GFAA) and inductively coupled plasma mass spectrometry (ICP-MS) will be compared and an overall approach to analysis described.
Learn more about our solutions: http://bit.ly/1cBJQDD
AOCS Plant Protein Science and Technology Forum ((https://plantprotein.aocs.org/) organized a series of virtual events during October 2020 to provide solution insight for the global protein challenge. Research Professor Nesli Sözer’s keynote presentation “Oats as an Alternative Protein Source” was part of the Plant Protein Science and Technology Forum's first session, "Processing and Utilization Technologies." The presentation's learning objectives were: opportunities and challenges of using oats as a protein source; fractionation and further modification technologies to improve oat protein functionality and oat protein-based meat and dairy alternative food examples.
This presentation describes a new food testing solution that allows mass detection to be accessible for many of the routine analyses found within a food testing lab. The ACQUITY UPLC, QDa detector in combination with the Empower software solution is fit for purpose and is easy to use. You simply power on and you are ready to go
With the ACQUITY QDa detector many of the normal processes that are required for mass spectrometers (such as mass calibration and optimisation and manual adjustments that need to be made) have all been fully automated.
In this application note, the amount of sugar or carbohydrate in a soft drink was determined using a colorimetric method. The rapid measurement of the PDA (Photodiode Array) UV/Vis Spectrophotometer allows for the collection of accurate data from the time-dependent reaction. The calibration curve was automatically calculated using the Quantification mode of the UV Lab™ software.
Accurate Determination of Lead in Different Dairy Products by Graphite Furnac...PerkinElmer, Inc.
This work describes a simple and
direct dilution method for sample preparation, followed by
automated analysis using GFAAS. This method minimizes
sample preparation, and also reduces potential contamination
while still maintaining the speed of analysis.
Learn more about our solutions: http://bit.ly/1bXfnRZ
2. 1. Introduction
Glucose
Glucose is by far the most important sub-
strate for microorganisms and mammalian
cell lines in bioprocesses. In over 90%
of all microbial cultivations and in all
mammalian cell cultivations it is used
as carbon source. Thus, monitoring and
control of glucose concentrations is
required. A typical example is a fed-batch
high cell density cultivation for the pro-
duction of recombinant proteins. The
concentration must remain below 1 g/L
during the production phase to minimize
the formation of unwanted byproducts.
Total depletion of glucose will immediately
lead to starvation and a decrease in cell
viability. Thus, the control of the substrate
feed requires frequent and reliable mea-
surements of the in-situ glucose concen-
tration.
Lactate
Lactate is a common metabolic product in
microbial and mammalian processes. The
monitoring and control of this metabolite
is necessary to avoid unwanted effects
when lactate concentration increases.
The productivity of mammalian cell cul-
tures is highly influenced by the lactate
concentration in the media. An increase
of the lactate concentration over a specific
value during the cultivation causes
negative cellular affects. Therefore, online
monitoring of the metabolite is necessary
for an optimal fed-batch process control.
In addition, the development of a process
where the final lactate concentration is
kept to a minimum, results in improved
purification procedures.
Analysis methods like HPLC or enzyme kits
are capable of measuring Glucose and
Lactate concentrations, but are relatively
expensive or problematic for online analysis.
BioPAT®
Trace and BioPAT®
Multi Trace
allows a rapid and precise determination
of Glucose and Lactate concentrations
inside the bioreactor within minutes.
3. System Performance
These data were compiled in order to give
an overview of the system- and sensor-
performance in the normal media concen-
tration range using the dialysis sampling
method.
Linearity
By comparing the actual value with the set
value a regression coefficient R2
of not less
than 0.9995 will be obtained (Figure 1).
0
2
4
6
8
10
12
14
16
18
20
0 2 4 6 8 10 12 14 16 18 20
Actualvalue[g/L]
Set value [g/L]
Linearity Glucose
y = 0,9707x + 0,0709
R² = 0,9998
0
1
2
3
4
5
6
7
8
9
10
11
0 1 2 3 4 5 6 7 8 9 10 11
y = 1,0165x + 0,0753
R² = 0,9996
Actualvalue[g/L]
Set value [g/L]
Linearity Glucose
Figure 1: Linearity of Glucose (R2
=0.9998) and
Lactate (R2
=0.9996)
2. Measurement Principle
Glucose
The enzyme glucose oxidase (GOD) is used
for the detection of glucose.
Glucose Gluconolactone
In presence of oxygen, glucose oxidase
catalyses the transformation of β-D-Glucose
to D-Glucono-δ-lactone and hydrogen
peroxide. The Glucose content is measured
indirectly via the formed peroxide, which is
oxidized to water and oxygen during the
amperometric measurement.
Lactate
The enzyme lactate oxidase (LOD) is used
for the detection of lactate.
L-Lactate Pyruvate
In presence of oxygen, lactate oxidase
catalyses the transformation of L-Lactate
to Pyruvat and hydrogen peroxide. The
Lactate content is measured indirectly via
the formed peroxide and the amperometric
measurement.
Amperometric measurement
The resulting hydrogen peroxide is oxidized
to water and oxygen generating a
measureable electric current which is
directly proportional to the parameter
concentration. This signal is based on a
calibrated linear range and gives the
in-situ concentration.
H2O2 → O2 + 2 H+
+ 2 e-
3. Precision
The typical variation about the mean value
is below 1.5% (Figure 2), except for the low
concentrations (< 2.5%).
0,0%
0,5%
1,0%
1,5%
2,0%
2,5%
3,0%
0,50 1,05 2,06 3,92 10,37 19,90
Variation[%]
Glucose [g/L]
Precision Glucose
0,0%
0,5%
1,0%
1,5%
2,0%
2,5%
3,0%
0,25 0,53 0,97 1,96 5,03 10,4
Precision Lactate
Lactate [g/L]
Variation[%]
Figure 2: Precision of Glucose and Lactate
Recovery
The recovery of the glucose and lactate
values is shown in figure 3.
0%
20%
40%
60%
80%
100%
120%
0,50 1,05 2,06 3,92 10,37 19,90
Recovery[%]
Glucose [g/L]
Recovery Glucose
0%
20%
40%
60%
80%
100%
120%
0,25 0,53 0,97 1,96 5,03 10,4
Recovery[%]
Lactate [g/L]
Recovery Lactate
Figure 3: Recovery of Glucose and Lactate
Operational stability
Long term stability for the application
Glucose|Lactate has been document tested
for 5,000 measurements or 14 days.
Operational stability
Long term stability for the application
Glucose|Lactate is guaranteed for 5,000
measurements or 14 days. Figure 4 shows a
typical profile during the load test within
the QA procedure over 5.000 assays (every
two minutes = 7 days).
0
20
40
60
80
100
120
0 1000 2000 3000 4000 5000
Glucose[%]
Measurement [number]
Long term stability: Glucose
0
20
40
60
80
100
120
0 1000 2000 3000 4000 5000
Lactate[%]
Measurement [number]
Long term stability: Lactate
Figure 4: Load test of Glucose and Lactate sensors
Shelf life
Glucose- | Lactate-Sensors have a shelf life
of at least 12 months at room temperature.
Consumables
Consumables for the application Glucose |
Lactate are listed in table 1:
Table 1: List of BioPAT®
Trace consumables
Article Article
number
Tubeset Dialysis BPT0003
Transport buffer 20x,
for cell cultivations
(Glucose | Lactate)
BPT0006
Transport buffer 5x,
for microbial cultivations
(Glucose | Lactate)
BPT0060
Membranes for Dialysis Probe;
5/pk
BPT0024
Calibration Standard 0.5 g/L
Glucose, 0.25 g/L Lactate
BPT0011
Calibration Standard 2 g/L
Glucose, 1 g/L Lactate
BPT0010
Calibration Standard 10 g/L
Glucose, 5 g/L Lactate
BPT0007
Cleaning solution BPT0044
4. Sales and Service Contacts
For further contacts, visit www.sartorius-stedim.com
Europe
Germany
Sartorius Stedim Biotech GmbH
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37079 Goettingen
Phone +49.551.308.0
Fax +49.551.308.3289
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Robert-Bosch-Strasse 5–7
34302 Guxhagen
Phone +49.5665.407.0
Fax +49.5665.407.2200
France
Sartorius Stedim FMT S.A.S.
ZI des Paluds
Avenue de Jouques – CS 91051
13781 Aubagne Cedex
Phone +33.442.845600
Fax +33.442.845619
Sartorius Stedim France SAS
ZI des Paluds
Avenue de Jouques – CS 71058
13781 Aubagne Cedex
Phone +33.442.845600
Fax +33.442.846545
Austria
Sartorius Stedim Austria GmbH
Modecenterstrasse 22
1030 Vienna
Phone +43.1.7965763.18
Fax +43.1.796576344
Belgium
Sartorius Stedim Belgium N.V.
Leuvensesteenweg, 248/B
1800 Vilvoorde
Phone +32.2.756.06.80
Fax +32.2.756.06.81
Hungary
Sartorius Stedim Hungária Kft.
Kagyló u. 5
2092 Budakeszi
Phone +36.23.457.227
Fax +36.23.457.147
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Sartorius Stedim Italy S.p.A.
Via dell’Antella, 76/A
50012 Antella-Bagno a Ripoli (FI)
Phone +39.055.63.40.41
Fax +39.055.63.40.526
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Sartorius Stedim Netherlands B.V.
Phone +31.30.60.25.080
Fax +31.30.60.25.099
filtratie.nederland@sartorius-stedim.com
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ul. Wrzesinska 70
62-025 Kostrzyn
Phone +48.61.647.38.40
Fax +48.61.879.25.04
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LLC “Sartorius ICR”and LLC “Biohit”
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Phone +7.812.327.5.327
Fax +7.812.327.5.323
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Edificio PAYMA
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Phone +34.902.110.935
Fax +34.91.358.96.23
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Phone +41.52.354.36.36
Fax +41.52.354.36.46
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Phone +44.1372.737159
Fax +44.1372.726171
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Post Box 440 “B”
01001 Kiev, Ukraine
Phone +380.44.411.4918
Fax +380.50.623.3162
America
USA
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Bohemia, NY 11716
Toll-Free +1.800.368.7178
Fax +1.631.254.4253
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Sartorius Argentina S.A.
Int. A. Ávalos 4251
B1605ECS Munro
Buenos Aires
Phone +54.11.4721.0505
Fax +54.11.4762.2333
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Avenida Senador Vergueiro 2962
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Phone +55.11.4362.8900
Fax + 55.11.4362.8901
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México
Phone +52.5555.62.1102
Fax +52.5555.62.2942
Asia|Pacific
Australia
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Phone +61.3.8762.1800
Fax +61.3.8762.1828
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Phone +86.10.80426516
Fax +86.10.80426580
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Trading Co. Ltd
3rd Floor, North Wing, Tower 1
No. 4560 Jinke Road
Zhangjiang Hi-Tech Park
Pudong District
Shanghai 201210, China
Phone +86.21.68782300
Fax +86.21.68782332|68782882
Sartorius Stedim Biotech (Beijing) Co. Ltd.
Guangzhou Representative Office
Unit K, Building 23
Huihua Commerce & Trade Building
No. 80 Xianlie Middle Road
Guangzhou 510070
Phone +86.20.37618687|37618651
Fax +86.20.37619051
India
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#69/2-69/3, NH 48, Jakkasandra
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Phone +91.80.4350.5250
Fax +91.80.4350.5253
Japan
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Phone +81.3.4331.4300
Fax +81.3.4331.4301
Malaysia
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Lot L3-E-3B, Enterprise 4
Technology Park Malaysia
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Phone +60.3.8996.0622
Fax +60.3.8996.0755
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The Capricorn, #05-08A,
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Singapore 117528
Phone +65.6872.3966
Fax +65.6778.2494
South Korea
Sartorius Korea Biotech Co., Ltd.
8th Floor, Solid Space B/D,
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Phone +82.31.622.5700
Fax +82.31.622.5799
www.sartorius-stedim.com
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PublicationNo.:SLL1104-e150401·OrderNo.:85037-550-18·Ver.04|2015